Sulfonamide-3,5-bis(benzylidene)-4-piperidinone derivatives, processes for their preparation and use

Abstract

The application discloses a series of sulfanilamide-3,5-bis(benzylidene)-4-piperidone derivatives, a preparation method and application thereof. The derivative is obtained by introducing sulfanilamide and valeramide on a 3,5-bis(benzylidene)-4-piperidone structure. Test results of the applicant show that part of the target compounds have good anti-tumor activity on breast cancer and colon cancer tumor cell lines, and are expected to be used for preparation of anti-tumor drugs.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical technology, specifically to sulfadiazine-3,5-bis(benzylidene)-4-piperidinone derivatives, their preparation methods, and applications. Background Technology

[0002] Carbonic anhydrase IX (CA IX) is a transmembrane, hypoxia-induced zinc metalloenzyme that converts carbon dioxide into bicarbonate and protons. This acid-base regulation function is crucial for maintaining an alkaline intracellular pH that promotes the survival and growth of malignant cells, as well as amplifying extracellular acidosis that promotes their invasion and metastasis. Cancer cells expressing CA IX often represent the most clinically aggressive component of heterogeneous tumors, making CA IX a key biomarker and a major therapeutic target. Carbonic anhydrase IX inhibitors mainly include sulfonamides and coumarins, with sulfonamides being the most studied. For example, compound E7070 (N-(3-chloro-7-indolyl)-1,4-benzenedisulfonamide) is a novel sulfonamide antitumor drug for the treatment of solid tumors, and it has entered phase II clinical trials (Indisulam: an anticancer sulfonamide in clinical development.[J]. Expert Opin Investig Drugs, 2003,12(2):283.). The mechanism of action of E7070 is to prevent the cell cycle from reaching the G1 phase, effectively inhibit carbonic anhydrase, and alter gene expression by nearly 60%. In 2016, Hai-Liang Zhu's research group reported a novel sulfonamide-substituted coumarin COX-2 inhibitor (Coumarinsulfonamides derivatives as potent and selective COX-2 inhibitors with efficacy in suppressing cancer proliferation and metastasis[J]. Bioorganic & Medicinal Chemistry Letters, 2016,26(15):3491-3498.). Among them, the sulfonamide-coumarin derivative with the best activity showed an activity of 360 nM against cancer cells HeLa and an inhibitory activity of 90 nM against COX-2. It could induce apoptosis in HeLa cells, and the apoptosis effect on HeLa cells was concentration-dependent and time-dependent.

[0003] Curcumin and its derivatives have attracted considerable attention over the past two decades due to their antitumor, antioxidant, and anti-inflammatory biological functions (Discovery of a new function of curcumin which enhances its anticancer therapeutic potency. Sci. Rep. 2016, 6, 30962.). The curcumin analog EF24 is a monocarbonyl curcumin analog with the following structure: Existing studies have shown that EF24 has good antitumor activity (EF24, a novel synthetic curcumin analog, induces apoptosis in cancer cells via a redox-dependent mechanism. Anticancer Drugs 2005;16:263–75. EF24 induces G2 / M arrest and apoptosis in cisplatin-resistant humanovarian cancer cells by increasing PTEN expression. J Biol Chem 2007;282:28609–18. etc.). The inventors of this application obtained a 3,5-bis(benzylidene)-4-piperidinone derivative with good antitumor activity (CN115260212A) by introducing biotin and polyethylene glycol into the structure of 3,5-bis(benzylidene)-4-piperidinone. However, there are currently no reports on the introduction of sulfonamides, valeramides, or other structures into the 3,5-bis(benzylidene)-4-piperidinone structure. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to provide a series of sulfadiazine-3,5-bis(benzylidene)-4-piperidinone derivatives with targeting properties and good activity against certain tumor cells, as well as their preparation methods and applications.

[0005] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:

[0006] The sulfonamide-3,5-bis(benzylidene)-4-piperidinone derivatives of this invention are compounds having the structure shown in formula (I) or formula (II) below, or pharmaceutically acceptable salts thereof:

[0007] (I) (II);

[0008] in:

[0009] R1 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group;

[0010] R2 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group;

[0011] R3 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group;

[0012] n represents 3.

[0013] Preferably, R1 represents a hydrogen atom, a halogen atom, a methyl atom, a methoxy atom, or a trifluoromethyl atom; R2 represents a hydrogen atom, a halogen atom, a methyl atom, a methoxy atom, or a trifluoromethyl atom; and R3 represents a hydrogen atom, a halogen atom, a methyl atom, a methoxy atom, or a trifluoromethyl atom.

[0014] The preparation method of the sulfonamide-3,5-bis(benzylidene)-4-piperidinone derivative of the present invention mainly includes the following steps: taking the compound shown in formula (III) below and p-aminobenzenesulfonamide or the compound shown in formula (IV) below in an organic solvent, reacting under heating or non-heating conditions to obtain the crude product of the compound shown in formula (I) or formula (II);

[0015] (III) (Ⅳ);

[0016] in:

[0017] R1 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group;

[0018] R2 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group;

[0019] R3 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group;

[0020] n represents 3.

[0021] The applicant discovered in experiments that adding a condensing agent during the reaction process can improve the yield of the target compound; therefore, it is preferable to add the condensing agent before the reaction. The condensing agent is a conventional choice or combination of conventional agents in the prior art, specifically selected from one or more combinations of HATU (2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate), DIPEA (N,N-diisopropylethylamine), HOBT (1-hydroxybenzotriazole), EDCI (1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride), and TBTU (O-benzotriazole-N,N,N',N'-tetramethylurea tetrafluoroboric acid), more preferably HATU, HOBT, or TBTU. The amount of condensing agent added is typically 1.2 to 2 times the molar amount of the compound shown in formula (III), preferably 1.5 to 2 times.

[0022] The applicant also discovered that carrying out the reaction under alkaline conditions can improve the yield of the target compound. Therefore, it is preferred that the reaction be carried out under alkaline conditions, and more preferably under alkaline conditions with a pH of 7.5 to 9. A conventional alkaline substance can be added to the system to make it alkaline. The alkaline substance can be a conventional inorganic base (such as potassium carbonate, sodium carbonate, or sodium bicarbonate), preferably potassium carbonate; or it can be an organic base (such as triethylamine, N,N-diisopropylethylamine, pyridine, or N-methylmorpholine), preferably triethylamine.

[0023] Since both an alkaline environment and the addition of a condensing agent are beneficial to increasing the yield of the target compound, the reaction in this application is more preferably carried out in the presence of a condensing agent and under alkaline conditions.

[0024] In the above preparation method, the organic solvent is preferably selected from one or more combinations of acetonitrile, acetone, methanol, ethanol, dichloromethane, chloroform, and tetrahydrofuran, and more preferably acetonitrile, methanol, or dichloromethane. The amount of organic solvent used is preferably sufficient to dissolve the reactants. Generally, based on 1 mmol of the compound represented by formula (III), all reactants are usually dissolved in 15 to 30 mL of organic solvent.

[0025] In the above preparation method, the reaction temperature can be between 20°C and the boiling point of the organic solvent, preferably 25-45°C. Higher reaction temperatures can accelerate the reaction rate. The reaction is monitored by TLC until complete. Based on the applicant's experience, when the reaction temperature is 25-45°C, a reaction time of 24-48 hours is suitable. After the reaction is complete, a precipitate will form in the resulting liquid. Collecting the precipitate yields the crude target compound. When a large amount of organic solvent is added initially, the resulting liquid can be concentrated and then allowed to stand overnight to precipitate more.

[0026] The crude product of the target compound obtained by the above method is therefore, the method of the present invention further includes a step of purifying the obtained crude product of the target compound. Specifically, conventional purification methods can be used to purify the crude product to improve the purity of each target compound, such as silica gel column chromatography. The eluent used for column chromatography is preferably a mixed solvent of dichloromethane and methanol. In the mixed solvent, the volume ratio of dichloromethane to methanol is preferably 50:1 to 50:8, more preferably 50:4 to 50:6.

[0027] In the preparation method described in this invention, the compound represented by formula (III) is obtained by a coordination reaction between compound S3 and compound S4 in an organic solvent:

[0028]

[0029] S3 S4;

[0030] The selection of R1, R2, R3, n, and organic solvents are as described above.

[0031] In preparing the compound shown in formula (III), to improve the yield, it is preferable to add a condensing agent before the reaction. The selection and amount of the condensing agent are the same as described above. The compound S3 involved can be prepared with reference to existing literature (Almansour AI, Kumar RS, Beevi F, et al. Facile, regio- and diastereoselective synthesis of spiro-pyrrolidine and pyrrolizine derivatives and evaluation of their antiproliferative activities[J]. Molecules (Basel, Switzerland), 2014,19(7):10033-10055.). The compound S4 involved is a C3~C6 fatty diacid, preferably glutaric acid. After the coordination reaction is completed, the obtained reaction material is concentrated and then an appropriate amount of ethyl acetate is added. A precipitate is formed. The precipitate is collected, which is the compound shown in formula (III). The obtained compound shown in formula (III) can be further purified using existing conventional techniques before being used in subsequent reactions.

[0032] In the preparation method described in this invention, the compound represented by formula (Ⅳ) is obtained by decarboxylation of acetazolamide (N-[5-(aminosulfonyl)-1,3,4-thiadiazol-2-yl]acetamide). Specifically, acetazolamide is placed in anhydrous ethanol under acidic conditions (pH=4~6) for decarboxylation. After the reaction is complete, the precipitate is collected, which is the compound represented by formula (Ⅳ). The obtained compound represented by formula (Ⅳ) can be further purified using existing conventional techniques before being used in subsequent reactions.

[0033] The applicant discovered through experiments that some derivatives of the present invention have good antitumor activity against a variety of tumor cell lines. Therefore, the present invention also includes the use of the above-mentioned sulfadiazine-3,5-bis(benzylidene)-4-piperidinone derivatives or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating tumors, and further in the preparation of medicaments for treating breast cancer or colon cancer.

[0034] Furthermore, the present invention also includes a pharmaceutical composition comprising a therapeutically effective dose of the above-mentioned sulfadiazine-3,5-bis(benzylidene)-4-piperidinone derivative or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier.

[0035] Compared with existing technologies, this invention provides a series of novel sulfadiazine-3,5-bis(benzylidene)-4-piperidinone derivatives and their preparation methods. The applicant's experimental results show that some of the target compounds of this invention exhibit good antitumor activity against breast cancer and colon cancer cell lines, and are expected to be used in the preparation of antitumor drugs. Attached Figure Description

[0036] Figure 1 The in vivo antitumor effects of compounds II10, II12, and II15 in the MDA-MB-231 xenograft tumor model in Experiment Example 3 are shown (mice were orally administered the carrier or compounds II10 (15 or 30 mg / kg), II12 (15 or 30 mg / kg), and II15 (15 or 30 mg / kg) every 3 days for 21 consecutive days). Among them, (A) is a photograph of tumor tissue isolated after 21 consecutive days of administration of compounds II10, II12, and II15; (B) is the weight of the tumor removed at the end of treatment with compounds II10, II12, and II15; (C) is the change in tumor volume in different treatment groups measured every 3 days; and (D) is the effect of compounds II10, II12, and II15 on the body weight of the MDA-MB-231 xenograft mouse model.

[0037] Figure 2 This is a single-crystal structure diagram of compound I3.

[0038] Figure 3 This is a single-crystal structure diagram of compound I14.

[0039] Figure 4 This is a single-crystal structure diagram of compound II3.

[0040] Figure 5 This is a single-crystal structure diagram of compound II10. Detailed Implementation

[0041] To better explain the technical solution of the present invention, the present invention will be further described in detail below with reference to the embodiments, but the implementation of the present invention is not limited thereto.

[0042] The compounds of formula (III) involved in the following examples were prepared according to the following synthetic route:

[0043]

[0044] R1 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; R2 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; R3 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; n represents 3.

[0045] The specific preparation method includes the following steps:

[0046] 1) Compound S1 (4-piperidinone, 10 mmol) and compound S2 (substituted benzaldehyde, 24 mmol) were placed in a round-bottom flask, glacial acetic acid (30 mL) was added, and the mixture was reacted under hydrogen chloride gas for 12 h. After the reaction was completed, the reaction mixture was filtered, and the filter cake was recrystallized with acetone (15 mL) and saturated sodium bicarbonate solution (15 mL), filtered, and compound S3 (yellow solid) was obtained.

[0047] 2) Place compound S3 (1 mmol) and compound S4 (1.2 mmol) in a round-bottom flask, add tetrahydrofuran (50 mL), 1.2 mmol triethylamine and 1.2 mmol HATU, heat to 40 °C and react for 3 h. After the reaction is complete, concentrate the obtained reaction material, and then add 2 mL ethyl acetate to the concentrate. A yellow solid precipitates out. Filter, collect the yellow solid, and dry to obtain the compound shown in compound formula (III).

[0048] The compounds of formula (Ⅳ) involved in the following examples were prepared according to the following synthetic route:

[0049]

[0050] (Ⅳ)

[0051] The specific preparation method is as follows: acetazolamide (22 mmol, purchased directly from the market) was placed in a round-bottom flask, anhydrous ethanol (30 mL) and hydrochloric acid (5 mL, 12 mol / L) were added, and the mixture was heated under reflux for 4 h. After the reaction was completed, the resulting reaction material was concentrated, and the concentrated solution was extracted with saturated sodium bicarbonate solution (15 mL) and ethyl acetate (20 mL). The ethyl acetate layer was collected, washed with saturated sodium chloride solution (10 mL), and concentrated. A precipitate was formed. The precipitate was dried to obtain the compound shown in formula (Ⅳ) (white solid).

[0052] Example 1

[0053] The sulfonamide-3,5-bis(benzylidene)-4-piperidinone derivative of the present invention with the structure shown in formula (I) was prepared according to the following synthetic route.

[0054]

[0055] I1: R1=F, R2=H, R3=H, n=3;

[0056] I2: R1=Cl, R2=H, R3=H, n=3;

[0057] I3: R1=CH3, R2=H, R3=H, n=3;

[0058] I4: R1=OCH3, R2=H, R3=H, n=3;

[0059] I5: R1=CF3, R2=H, R3=H, n=3;

[0060] I6: R1=H, R2=F, R3=H, n=3;

[0061] I7: R1=H, R2=Cl, R3=H, n=3;

[0062] I8: R1=H, R2=CH3, R3=H, n=3;

[0063] I9: R1=H, R2=OCH3, R3=H, n=3;

[0064] I10: R1=H, R2=CF3, R3=H, n=3;

[0065] I11: R1=H, R2=H, R3=F, n=3;

[0066] I12: R1=H, R2=H, R3=Cl, n=3;

[0067] I13: R1=H, R2=H, R3=CH3, n=3;

[0068] I14: R1=H, R2=H, R3=OCH3, n=3;

[0069] I15: R1=H, R2=H, R3=CF3, n=3.

[0070] The specific preparation method is as follows: In a round-bottom flask, add 1 mmol of the compound shown in formula (III), 1 mmol of p-aminobenzenesulfonamide, 1.2 mmol of triethylamine, 1.2 mmol of HATU, and 20 mL of dichloromethane. After stirring until homogeneous, the pH of the system is 7.5-9. The reaction is carried out at room temperature for 24-48 h (TLC monitoring of the reaction). After the reaction is complete, the reactants are mixed and purified by silica gel column chromatography (dichloromethane / methanol = 50 / 6, volume ratio) to obtain target compound I. Different target products and their characterization are as follows:

[0071] 5-(3,5-bis((E)-2-fluorobenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I1): Yield 62%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ 10.10 (s,1H), 7.76 – 7.71 (m, 4H), 7.67 (d, J = 8.9Hz, 2H), 7.54 (dq, J = 18.9, 10.9,9.1 Hz, 4H), 7.39 – 7.29 (m, 4H), 7.24 (s, 2H), 4.75 (s, 4H), 2.23 (q, J =6.9 Hz, 4H), 1.66 (p, J =7.2Hz, 2H). 13 C NMR (100MHz, DMSO-d6) δ 185.62,171.19, 170.72, 142.08,138.01,131.99, 131.91, 130.97, 130.95, 128.37, 126.54,124.76, 124.73, 118.42, 115.95, 115.78, 35.20, 30.97, 20.00 ppm. HR–MS(m / z)(ESI): calcd for C 30 H27F2N3O5S [M+H] + : 580.1710; found: 580.1639.

[0072] 5-(3,5-bis((E)-2-chlorobenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I2): Yield 64%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ10.11 (s, 1H), 7.82 (d, J = 26.6 Hz, 2H), 7.75 – 7.70 (m, 2H), 7.69 – 7.65(m, 2H), 7.64 – 7.55 (m, 3H), 7.52 – 7.43 (m, 5H), 7.24 (s, 2H), 4.71 (d, J =20.7 Hz, 4H), 2.19 (dt, J = 27.5, 7.2 Hz, 4H), 1.65 (p, J = 7.2 Hz, 2H). 13 CNMR (100 MHz, DMSO-d6) δ 186.12, 171.15, 170.50, 142.08, 138.03, 135.17,135.11, 130.40, 129.33, 128.95, 126.56, 125.86, 118.43, 45.77, 42.56, 35.19,30.95, 19.92, 19.57 ppm. HR–MS (m / z)(ESI): calcd for C 30 H27F2N3O5S [M+Na] + :634.0951; found: 634.1048.

[0073] 5-(3,5-Di((E)-2-methylbenzylmethylene)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I3): Yield 65%, as a yellow solid. ¹H NMR (500 MHz, DMSO-d6) δ 10.10 (s, 1H), 7.84 (d, J = 21.0 Hz, 2H), 7.76 – 7.66 (m, 4H), 7.35 – 7.24 (m, 10H), 4.65 (d, J = 36.3 Hz, 4H), 2.30 (d, J = 21.2 Hz, 6H), 2.19 (t, J = 7.3 Hz, 2H), 2.10 (t, J = 7.3 Hz, 2H), 1.62 (t, J = 7.2Hz, 2H).13 C NMR (100 MHz, DMSO-d6) δ 186.17, 171.22, 170.55, 142.14, 138.06, 137.92, 137.80, 135.17,133.37, 133.18, 132.88, 132.82, 130.47, 130.38, 129.42, 129.04, 126.63,125.92, 118.47, 45.83, 42.63, 35.22, 30.99, 19.95, 19.65 ppm. HR–MS(m / z)(ESI): calcd for C 32 H 33 N3O5S [M+H] + : 572.2215; found: 572.2141.

[0074] 5-(3,5-bis((E)-2-methoxybenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I4): Yield 66%, as a yellow solid. 1H NMR (500 MHz, DMSO-d6) δ10.11 (s, 1H), 7.89 (s, 1H), 7.84 (s, 1H), 7.73 (d, J = 8.9 Hz, 2H), 7.68 (d,J = 8.9 Hz, 2H), 7.48 – 7.41 (m, 2H), 7.36 (dd, J = 14.2, 7.5 Hz, 2H), 7.24(s, 2H), 7.07 (ddd, J = 33.1, 17.0, 8.0 Hz, 4H), 4.72 (d, J = 17.3 Hz, 4H), 3.85 (d, J = 14.3 Hz, 6H), 2.25 – 2.15(m, 4H), 1.65 (p, J = 7.3 Hz, 2H).13CNMR (100 MHz, DMSO-d6) δ 185.61, 171.19, 170.71, 142.08, 138.01, 131.98,131.91, 130.97, 130.94, 128.37, 126.54, 124.76, 124.72, 118.42, 115.95,115.77, 46.08, 42.41, 35.20, 30.97, 20.00 ppm. HR–MS(m / z)(ESI): calcd forC 32 H 33 N3O7S[M+H]+: 604.2121; found: 604.2039.

[0075] 5-(3,5-bis((E)-2-trifluoromethylbenzylmethylene)-4-oxoperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I5): Yield 67%, as a yellow solid. 1 H NMR(400MHz, DMSO-d6) δ10.09 (s, 1H), 7.88 (d, J = 7.6 Hz, 3H), 7.81 (t, J = 7.9 Hz, 2H), 7.78 –7.70 (m, 3H), 7.70 – 7.53 (m, 6H), 7.25 (s, 2H), 4.62 (d, J = 18.3 Hz, 4H), 2.19 (t, J = 7.2 Hz, 2H), 2.10 (t, J = 7.3 Hz, 2H), 1.62 (p, J = 7.3 Hz, 2H).13 C NMR (100 MHz, DMSO-d6) δ 185.99, 171.59, 171.08, 142.56, 138.47, 135.09,133.21, 133.03, 132.88, 132.68, 131.30, 130.14, 127.05, 126.86, 126.81,125.81, 123.09, 118.88, 46.12, 42.74, 35.63, 31.31, 20.36 ppm. HR–MS(m / z)(ESI): calcd for C 32 H 27 F6N3O5S[M+H] + : 680.1642; found: 680.1576.

[0076] 5-(3,5-bis((E)-3-fluorobenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I6): Yield 66%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ 10.10(s, 1H), 7.74 – 7.71 (m, 2H), 7.70 – 7.63 (m, 4H), 7.58 – 7.48 (m, 2H), 7.41(td, J = 14.7, 12.7, 7.9 Hz, 4H), 7.30 (d, J = 9.4 Hz, 2H), 7.22 (s, 2H), 4.84 (s, 4H), 2.26 (dt, J = 9.4, 7.2 Hz, 4H), 1.68 (p, J = 7.2 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ186.09,171.20,170.77,163.05,161.10,142.06,134.95,134.84,130.77,130.69,126.53,118.41,46.07, 42.28, 35.20, 31.03, 20.05 ppm. HR–MS (m / z)(ESI): calcd for C 30 H 27 F2N3O5S [M+H] + : 580.1710; found: 580.1639.

[0077] 5-(3,5-bis((E)-3-chlorobenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I7): Yield 67%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ 10.12(s, 1H), 7.73 – 7.61 (m, 8H), 7.51 (d, J = 20.4 Hz, 6H), 7.24 (s, 2H), 4.83(s, 4H), 2.28 – 2.22 (m, 4H), 1.71 – 1.64 (m, 2H). 13 C NMR (100MHz, DMSO-d6) δ186.12, 171.15, 170.50, 142.08, 138.03, 135.17, 135.11, 130.40, 129.33,128.95, 126.56, 125.86, 118.43, 45.77, 42.56, 35.19, 30.95, 19.92, 19.57 ppm.HR–MS (m / z) (ESI): calcd for C 30 H 27 F2N3O5S [M + Na] + : 634.0951; found: 634.1048.

[0078] 5-(3,5-bis((E)-3-methylbenzylmethylene)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I8): Yield 68%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ 10.11(s, 1H), 7.76 – 7.72 (m, 2H), 7.72 – 7.63 (m, 4H), 7.44 (dd, J = 13.2, 7.8Hz, 4H), 7.33 – 7.23 (m, 6H), 4.82 (d, J = 9.7 Hz, 4H), 2.35 (d, J = 10.9 Hz, 6H), 2.24 (t, J = 7.2 Hz, 4H), 1.68 (p, J = 7.2 Hz, 2H). 13C NMR (100MHz, DMSO-d6) δ 186.16, 171.36, 170.73, 142.18, 138.22, 138.17, 138.08, 136.48, 136.38,134.42, 134.12, 132.63, 132.44, 131.22, 131.13, 130.43, 130.37, 128.80,127.69, 127.57, 126.66, 118.53, 46.26, 42.68, 35.31, 31.10, 21.03, 20.15ppm. (m / z) (ESI): calcd for C 32 H 33 N3O5S [M + H] + : 572.2215; found: 572.2141.

[0079] 5-(3,5-bis((E)-3-methoxybenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I9): Yield 68%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ 10.10(s, 1H), 7.75 – 7.71 (m, 2H), 7.70 – 7.64 (m, 4H), 7.41 (dt, J = 20.1, 8.0Hz, 2H), 7.22 (s, 2H), 7.10 (d, J = 12.5 Hz, 4H), 7.06 – 6.99 (m, 2H), 4.84(s, 4H), 3.80 (d, J = 9.7 Hz, 6H), 2.25 (q, J =7.3Hz, 4H), 1.68 (p, J = 7.2Hz, 2H). 13 C NMR (100MHz, DMSO-d6) δ 186.13, 171.22, 170.66, 159.33, 142.07,138.01, 136.15, 129.85, 126.54, 122.58, 118.44, 115.74, 115.61, 115.52,115.31, 55.21, 46.13, 42.44, 35.24, 31.05, 20.06 ppm. HR–MS (m / z) (ESI):calcd for C 32H 33 N3O7S[M + H] + : 604.2121; found: 604.2039.

[0080] 5-(3,5-bis((E)-3-trifluoromethylbenzylmethylene)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I10): Yield 67%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ10.09 (s, 1H), 7.91 (d, J = 5.0 Hz, 2H), 7.88 – 7.78 (m, 5H), 7.76 (s, 2H),7.74 – 7.69 (m, 3H), 7.68 – 7.64 (m, 2H), 7.22 (s, 2H), 4.85 (d, J = 8.9 Hz, 4H), 2.25 (q, J = 7.5 Hz, 4H), 1.67 (p, J = 7.2Hz, 2H). 13 C NMR (100MHz, DMSO-d6) δ 186.04, 171.17, 170.78, 142.05, 138.00, 134.74, 134.58, 133.86, 133.74,129.85, 126.52, 118.38, 45.99, 42.11, 35.17, 31.04, 20.01 ppm. HR–MS (m / z)(ESI): calcd for C 32 H 27 F6N3O5S[M + H] + : 680.1642; found: 680.1576.

[0081] 5-(3,5-bis((E)-4-fluorobenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I11): Yield 70%, as a yellow solid. 1H NMR (500MHz, DMSO-d6) δ 10.10(s, 1H), 7.75 – 7.71 (m, 2H), 7.71 – 7.65 (m, 4H), 7.65 – 7.58 (m, 4H), 7.32(dt, J = 24.0, 8.6 Hz, 4H), 7.23 (s, 2H), 4.82 (s, 4H), 2.25 (td, J = 7.2,2.5 Hz, 4H), 1.68 (p, J = 7.2 Hz, 2H). 13 C NMR (100MHz, DMSO-d6) δ 186.01,171.20, 170.68, 163.51, 161.53, 142.06, 138.02, 135.02, 132.95, 132.88,132.81, 126.54, 118.41, 115.92, 115.75, 46.02, 42.35, 35.20, 31.02, 20.03ppm. HR–MS(m / z) (ESI): calcd for C 30 H 27 F2N3O5S [M + H] + : 580.1710; found:580.1639.

[0082] 5-(3,5-bis((E)-4-chlorobenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I12): Yield 64%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ 10.10(s, 1H), 7.75 – 7.72 (m, 2H), 7.69 – 7.63 (m, 4H), 7.60 – 7.49 (m, 8H), 7.22(s, 2H), 4.82 (s, 4H), 2.30 – 2.20 (m, 4H), 1.68 (p, J = 7.1 Hz, 2H). 13C NMR(100MHz, DMSO-d6) δ 185.94,171.19,170.01,142.06,138.01, 134.86, 132.21,128.82, 126.55, 118.40, 46.04,42.38,35.19, 31.02, 20.01 ppm. HR–MS (m / z)(ESI): calcd for C 30 H 27 F2N3O5S [M + Na] + : 634.0951; found: 634.1048.

[0083] 5-(3,5-bis((E)-4-methylbenzylmethylene)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I13): Yield 69%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ10.11 (s, 1H), 7.75 – 7.72 (m, 2H), 7.71 – 7.63 (m, 4H), 7.44 (dd, J = 13.2,7.8 Hz, 4H), 7.34 – 7.24 (m, 6H), 4.82 (d, J = 9.7 Hz, 4H), 2.35 (d, J =10.9Hz, 6H), 2.24 (t, J = 7.2 Hz, 4H), 1.68 (p, J = 7.2 Hz, 2H). 13 C NMR(100MHz, DMSO-d6) δ 186.16, 171.36, 170.73, 142.18, 138.22, 138.17, 138.08,136.48, 136.38, 134.42, 134.12, 132.63, 132.44, 131.22, 131.13, 130.43,130.37, 128.80, 127.69, 127.57, 126.66, 118.53, 46.26, 42.68, 35.31, 31.10,21.03, 20.15 ppm. HR–MS (m / z) (ESI): calcd for C 32 H 33 N3O5S [M + H] + : 572.2215; found: 572.2141.

[0084] 5-(3,5-bis((E)-4-methoxybenzylmethyl)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I14): Yield 63%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6) δ10.10 (s, 1H), 7.75 – 7.71 (m, 2H), 7.69 – 7.61 (m, 4H), 7.52 (t, J = 8.0 Hz,4H), 7.22 (s, 2H), 7.05 (dd, J = 20.9, 8.4 Hz, 4H), 4.83 (d, J = 5.9 Hz, 4H), 3.82 (d, J = 11.6 Hz, 6H), 2.32 – 2.24 (m, 4H), 1.70 (p, J = 7.2 Hz, 2H). 13 CNMR (100 MHz, DMSO-d6) δ 160.31, 135.81, 135.75, 132.54, 132.48, 126.54,118.43, 114.36, 55.31, 35.23, 31.05, 20.08 ppm. HR–MS(m / z)(ESI): calcd forC 32 H 33 N3O7S[M + H] + : 604.2121; found: 604.2039.

[0085] 5-(3,5-bis((E)-4-trifluoromethylbenzylmethylene)-4-oxopiperidin-1-yl)-5-oxo-N-(4-sulfonylphenyl)pentanamide (I15): Yield 69%, as a yellow solid. 1 H NMR (500MHz, DMSO-d6)δ 10.09 (s, 1H), 7.84 (dd, J = 20.6, 8.0 Hz, 4H), 7.79 – 7.71 (m, 8H), 7.68 –7.65 (m, 2H), 7.22 (s, 2H), 4.85 (s, 4H), 2.28 – 2.22 (m, 4H), 1.68 (p, J =7.2 Hz, 2H). 13C NMR (100 MHz, DMSO-d6) δ 186.06, 171.21, 170.78, 142.06,138.03, 134.56, 131.02, 130.92, 126.53, 125.55, 125.52, 125.49, 125.45,118.38, 46.04, 42.29, 35.19, 30.97, 19.97 ppm.HR–MS (m / z)(ESI): calcd forC 32 H 27 F6N3O5S[M + H] + : 680.1642; found: 680.1576.

[0086] The structural formulas of the target compounds I1~I15 obtained in this embodiment are shown below:

[0087]

[0088] Example 2: Preparation of target compounds I1 and I2

[0089] Compound I1: Example 1 was repeated, except that triethylamine was not added. A yellow solid was obtained in 42% yield. Characterization by 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed it to be compound I1.

[0090] Compound I1: Example 1 was repeated, except that triethylamine and HATU were not added. A yellow solid was obtained in 33% yield. Characterization by 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed it to be compound I1.

[0091] Compound I2: Example 1 was repeated, except that acetonitrile was used instead of dichloromethane, and the reaction was carried out at 35°C. A yellow solid was finally obtained in 60% yield. Characterization by 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed it to be compound I2.

[0092] Example 3: Preparation of single crystals of target compounds I3 and I14

[0093] Compound I3 Single Crystal: 10 mg of Compound I3 prepared in Example 1 was placed in a beaker, and 10 ml of a mixture of acetonitrile and dichloromethane (volume ratio 1:1) was added. Then, 40 ml of water was added, the beaker was sealed with sealing film, and placed in a cool place for slow evaporation for 5 days. Pale yellow triclinic columnar crystals precipitated. Crystals of suitable size were selected for structural analysis using X-ray single-crystal diffraction. Detailed crystal measurement data are shown in Table 1 below. The crystal structure of the obtained crystals is as follows: Figure 2 As shown, the obtained pale yellow triclinic columnar crystals were determined to be single crystals of the target compound I3.

[0094] Compound I14 Single Crystal: 10 mg of compound I14 prepared in Example 1 was placed in a beaker, and 10 ml of a mixture of acetonitrile and dichloromethane (volume ratio 1:1) was added. Then, 40 ml of water was added, the beaker was sealed with sealing film, and placed in a cool place for slow evaporation for 5 days. Pale yellow triclinic columnar crystals precipitated. Crystals of suitable size were selected for structural analysis using X-ray single-crystal diffraction. Detailed crystal measurement data are shown in Table 1 below. The crystal structure of the obtained crystals is as follows. Figure 3 As shown, the obtained pale yellow triclinic columnar crystals were determined to be single crystals of the target compound I14.

[0095] Table 1: Data on crystallographic and structural corrections for compounds I3 and I14

[0096]

[0097] Example 4

[0098] The sulfonamide-3,5-bis(benzylidene)-4-piperidinone derivatives of the structure shown in formula (II) of the present invention were prepared according to the following synthetic route.

[0099]

[0100] Ⅱ1: R1=F, R2=H, R3=H, n=3;

[0101] Ⅱ2: R1=Cl, R2=H, R3=H, n=3;

[0102] Ⅱ3: R1=CH3, R2=H, R3=H, n=3;

[0103] Ⅱ4: R1=OCH3, R2=H, R3=H, n=3;

[0104] Ⅱ5: R1=CF3, R2=H, R3=H, n=3;

[0105] Ⅱ6: R1=H, R2=F, R3=H, n=3;

[0106] Ⅱ7: R1=H, R2=Cl, R3=H, n=3;

[0107] Ⅱ8: R1=H, R2=CH3, R3=H, n=3;

[0108] Ⅱ9: R1=H, R2=OCH3, R3=H, n=3;

[0109] Ⅱ10: R1=H, R2=CF3, R3=H, n=3;

[0110] Ⅱ11: R1=H, R2=H, R3=F, n=3;

[0111] Ⅱ12: R1=H, R2=H, R3=Cl, n=3;

[0112] Ⅱ13: R1=H, R2=H, R3=CH3, n=3;

[0113] Ⅱ14: R1=H, R2=H, R3=OCH3, n=3;

[0114] Ⅱ15: R1=H, R2=H, R3=CF3, n=3.

[0115] The specific preparation method is as follows: In a round-bottom flask, add 1 mmol of the compound shown in formula (III), 1 mmol of the compound shown in formula (IV), 1.2 mmol of triethylamine, 1.2 mmol of HATU, and 20 mL of dichloromethane. After stirring until homogeneous, the pH of the system is 7.5-9. The reaction is carried out at room temperature with stirring for 24-48 h (TLC monitoring of the reaction). After the reaction is complete, the reactants are mixed and purified by silica gel column chromatography (dichloromethane / methanol = 50 / 6, volume ratio) to obtain target compound II. Different target products and their characterization are as follows:

[0116] 5-(3,5-bis((E)-2-fluorobenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanilamide-1,3,4-thiadiazol-2-yl)pentanamide (II1): Yield 67%, as a yellow solid. 1 H NMR (500MHz, DMSO) δ 12.74 (s, 1H), 8.31 (s, 2H), 7.73 (d, J = 15.8 Hz, 2H), 7.58 – 7.49(m, 4H), 7.33 (dd, J = 13.5, 7.9 Hz, 4H), 4.74 (d, J = 8.5 Hz, 4H), 2.42 (t,J = 7.2 Hz, 2H), 2.23 (t, J = 7.2 Hz, 2H), 1.70 (p, J = 7.2 Hz, 2H). 13C NMR(100 MHz, DMSO-d6) δ 185.61, 171.69, 170.51, 164.27, 163.92, 160.94, 154.12,134.23, 131.93, 130.92, 128.39, 124.72, 115.93, 115.76, 46.01, 42.39, 33.77,30.70, 19.41 ppm.HR–MS (m / z)(ESI): calcd for C 26 H 22 F2N5O5S2[M+H] + : 588.1007; found: 588.1109.

[0117] 5-(3,5-bis((E)-2-chlorobenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ2): Yield 68%, as a yellow solid. 1 H NMR(500MHz,DMSO) δ 12.86 (s, 1H), 8.32 (s, 2H), 7.81 (d, J = 29.3 Hz, 2H), 7.65 – 7.39(m, 8H), 4.69 (d, J = 26.4 Hz, 4H), 2.42 (t, J = 7.2 Hz, 2H), 2.17 (t, J =7.2 Hz, 2H), 1.69 (p, J = 7.2 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 185.88,171.64, 170.47, 164.26, 160.95, 134.04, 133.83, 132.76, 132.16, 131.09,130.81, 129.79, 127.35, 45.68, 42.23, 33.78, 30.71, 19.37 ppm.HR–MS(m / z)(ESI): calcd for C 26 H 23 Cl2N5O5S2[M+H] + : 620.0591; found: 620.0518.

[0118] 5-(3,5-bis((E)-2-methylbenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (II3): Yield 69%, as a yellow solid. 1 H NMR(500MHz,DMSO) δ 12.85 (s, 1H), 8.32 (s, 2H), 7.83 (d, J = 20.0 Hz, 2H), 7.37 – 7.22(m, 8H), 4.64 (d, J = 37.7 Hz, 4H), 2.38 (t, J = 7.2 Hz, 2H), 2.30 (d, J =15.6 Hz, 6H), 2.10 (t, J = 7.2 Hz, 2H), 1.65 (p, J = 7.2 Hz, 2H). 13 C NMR(100MHz, DMSO-d6) δ 186.09, 171.65, 170.26, 164.27, 160.93, 137.70, 135.11,133.23, 133.10 – 133.01, 132.80, 130.31, 129.29, 128.91, 125.83, 45.69,42.53, 33.74, 30.70,19.52,19.34 ppm.HR–MS(m / z)(ESI): calcd for C 28 H 29 N5O5S2[M+H] + : 580.1693; found: 580.1610.

[0119] 5-(3,5-bis((E)-2-methoxybenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (II4): Yield 63%, as a yellow solid. 1H NMR(500MHz,DMSO) δ 12.85 (s, 1H), 8.32 (s, 2H), 7.85 (d, J = 29.7 Hz, 2H), 7.48 – 7.31(m, 4H), 7.07 (m, J = 22.0, 17.3, 7.9 Hz, 4H), 4.70 (d, J = 21.4 Hz, 4H), 3.85 (d, J = 8.1 Hz, 6H), 2.40 (t, J = 7.2 Hz, 2H), 2.19 (t, J = 7.2 Hz, 2H), 1.68 (p, J = 7.2 Hz, 2H). 13 C NMR (100MHz, DMSO-d6) δ 186.20, 171.71, 170.30,164.26, 160.95, 158.02, 132.25, 131.70, 131.36, 130.18, 122.88, 122.65,120.31 , 111.38 , 55.59, 46.08 , 42.55, 33.83, 30.81, 19.47, 18.62 ppm.HR–MS(m / z)(ESI): calcd for C 28 H 29 N5O7S2[M+H] + : 612.1587; found: 612.1508.

[0120] 5-(3,5-bis((E)-2-trifluoromethylbenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ5): Yield 65%, as a yellow solid. 1 H NMR (500MHz, DMSO) δ 12.86 (s, 1H), 8.32 (s, 2H), 7.93 – 7.75 (m, 6H), 7.69 – 7.51(m, 4H), 4.61 (d, J = 27.5 Hz, 4H), 2.39 (t, J = 7.2 Hz, 2H), 2.12 (t, J =7.2 Hz, 2H), 1.66 (p, J = 7.2 Hz, 2H). 13C NMR(100MHz, DMSO-d6) δ 185.48,171.62, 170.36, 164.27, 160.93, 134.58, 132.62, 130.73, 129.59, 126.31,125.02, 122.84, 45.55, 42.20,33.76,30.5,19.33ppm.HR–MS(m / z)(ESI):calcd forC 28 H 23 F6N5O5S2[M+H] + : 688.1122; found: 688.1045.

[0121] 5-(3,5-bis((E)-3-fluorobenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanilamide-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ6): Yield 68%, as a yellow solid. 1 H NMR(500MHz, DMSO)δ 12.87 (s, 1H), 8.32 (s, 2H), 7.67 (d, J = 14.2 Hz, 2H), 7.58 – 7.48(m, 2H), 7.45 – 7.36 (m, 4H), 7.33 – 7.25 (m, 2H), 4.83 (s, 4H), 2.44 (t, J =7.2 Hz, 2H), 2.28 (t, J = 7.2 Hz, 2H), 1.76 – 1.66 (m, 2H). 13 C NMR (100MHz, DMSO-d6) δ 186.11, 171.74, 170.59, 164.26, 163.05, 161.04, 136.48, 134.91,133.60, 130.75, 126.54, 116.98, 46.01, 42.27, 33.81, 30.81, 19.45 ppm.HR–MS (m / z) (ESI): calcd for C 26 H 22 F2N5O5S2[M+H] + : 588.1007; found: 588.1109.

[0122] 5-(3,5-bis((E)-3-chlorobenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ7): Yield 65%, as a yellow solid. 1 H NMR (500MHz, DMSO) δ 12.88 (s, 1H), 8.31 (s, 2H), 7.64 (dd, J = 17.0, 10.5 Hz, 4H), 7.51 (d, J = 14.1 Hz, 6H), 4.81 (s, 4H), 2.44 (t, J = 7.2 Hz, 2H), 2.27 (t, J =7.2 Hz, 2H), 1.70 (p, J = 7.3 Hz, 2H). 13 C NMR (100MHz, DMSO-d6) δ 86.02,171.74, 170.58, 164.25, 161.00, 136.41, 136.16, 134.74, 133.74, 133.52,130.56, 129.91, 129.29, 128.76, 45.97, 42.24, 33.81, 30.80, 19.44 ppm.HR–MS(m / z) (ESI): calcd for C 26 H 23 Cl2N5O5S2[M+H] + : 620.0591; found: 620.0518.

[0123] 5-(3,5-bis((E)-3-methylbenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (II8): Yield 64%, as a yellow solid. 1H NMR (500MHz, DMSO) δ 12.87 (s, 1H), 8.33 (s, 2H), 7.66 (d, J = 12.8 Hz, 2H), 7.36 (dd, J =24.8, 8.5 Hz, 6H), 7.26 (dd, J = 16.0, 7.4 Hz, 2H), 4.82 (d, J = 12.2 Hz, 4H), 2.42 (t, J = 7.2 Hz, 2H), 2.36 (d, J = 11.0 Hz, 6H), 2.24 (t, J = 7.2Hz, 2H), 1.70 (p, J = 7.1 Hz, 2H). 13 C NMR(100 MHz, DMSO-d6) δ 186.08, 171.72,170.45, 164.27, 160.97, 138.08, 136.30, 134.33, 134.05, 132.46, 131.03,130.28, 128.69, 127.49, 46.10, 42.58, 33.81, 30.82, 20.93, 19.46 ppm.HR–MS(m / z)(ESI): calcd for C 28 H 29 N5O5S2[M+H] + : 580.1693; found: 580.1610.

[0124] 5-(3,5-bis((E)-3-methoxybenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ9): Yield 64%, as a yellow solid. 1 H NMR (500MHz, DMSO) δ 12.87 (s, 1H), 8.32 (s, 2H), 7.67 (d, J = 12.4 Hz, 2H), 7.41 (dt, J =17.0, 8.0 Hz, 2H), 7.13 – 7.07 (m, 4H), 7.06 – 6.99 (m, 2H), 4.83 (d, J = 6.4Hz, 4H), 3.80 (d, J = 8.9 Hz, 6H), 2.43 (t, J = 7.3 Hz, 2H), 2.27 (t, J = 7.2Hz, 2H), 1.71 (p, J = 7.3 Hz, 2H).13 C NMR (100MHz, DMSO-d6) δ 186.14, 171.73,170.49, 164.26, 160.98, 159.33, 136.17, 135.68, 135.43, 132.78, 129.86,122.56, 115.69, 115.39, 55.20, 46.07, 42.42, 33.81, 30.84, 19.45 ppm.HR–MS(m / z) (ESI): calcd for C 28 H 29 N5O7S2[M+H] + : 612.1587; found: 612.1508.

[0125] 5-(3,5-bis((E)-3-trifluoromethylbenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (II10): Yield 67%, as a yellow solid. 1 H NMR(500MHz, DMSO) δ 12.88 (s, 1H), 8.31 (s, 2H), 7.96 – 7.68 (m, 10H), 4.84 (d,J = 2.8 Hz, 4H), 2.43 (t, J = 7.3 Hz, 2H), 2.27 (t, J = 7.2 Hz, 2H), 1.70 (p,J = 7.2 Hz, 2H). 13 C NMR(100MHz, DMSO-d6) δ 186.08, 171.74, 170.61, 164.24,161.00, 135.24, 134.68, 134.05, 133.80, 129.80, 129.48, 127.11, 126.82,125.87, 125.00, 122.84, 45.94, 42.08, 33.81, 30.83, 19.44 ppm.HR–MS(m / z)(ESI): calcd for C 28 H 23 F6N5O5S2[M+H] + : 688.1122; found: 688.1045.

[0126] 5-(3,5-bis((E)-4-fluorobenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ11): Yield 63%, as a yellow solid. 1 H NMR (500MHz, DMSO) δ 12.89 (s, 1H), 8.33 (s, 2H), 7.72 – 7.57 (m, 6H), 7.32 (dt, J = 21.6,8.6 Hz, 4H), 4.79 (t, J = 9.3 Hz, 4H), 2.43 (t, J = 7.2 Hz, 2H), 2.26 (t, J =7.1 Hz, 2H), 1.71 (p, J = 7.2 Hz, 2H). 13 C NMR(100MHz, DMSO-d6) δ 186.08,171.76, 170.51, 164.26, 163.53, 161.55, 160.98, 135.04, 132.90, 132.35,130.92, 130.67, 115.96, 115.79, 45.96, 42.32, 33.81, 30.79, 19.43 ppm.HR–MS(m / z)(ESI): calcd for C 26 H 22 F2N5O5S2[M+H] + : 588.1007; found: 588.1109.

[0127] 5-(3,5-bis((E)-4-chlorobenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ12): Yield 69%, as a yellow solid. 1 H NMR (500 MHz, DMSO) δ 12.88 (s, 1H), 8.32 (s, 2H), 7.66 (d, J = 12.9 Hz, 2H), 7.54 (dd, J =18.9, 8.4 Hz, 8H), 4.80 (s, 4H), 2.43 (t, J = 7.2 Hz, 2H), 2.26 (t, J =7.1Hz, 2H), 1.70 (p, J =7.2Hz, 2H). 13C NMR (100MHz, DMSO-d6) δ 185.99, 171.73,170.52, 164.25, 160.96, 134.86, 134.30, 133.46 – 132.77, 132.17, 128.83,45.94, 42.33, 33.80, 30.75, 19.38 ppm.HR–MS (m / z)(ESI): calcd forC 26 H 23 Cl2N5O5S2[M+H] + : 620.0591; found: 620.0518.

[0128] 5-(3,5-bis((E)-4-methylbenzyl)-4-oxoperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ13): Yield 65%, as a yellow solid. 1 H NMR (500MHz, DMSO) δ 12.86 (s, 1H), 8.32 (s, 2H), 7.65 (d, J = 13.3 Hz, 2H), 7.43 (dd, J =11.9, 8.1 Hz, 4H), 7.29 (dd, J = 22.5, 7.7 Hz, 13C NMR (100 MHz, DMSO-d6) δ 186.00,171.78, 170.40, 164.21, 161.05, 139.55, 136.09, 131.80, 131.47, 131.32 –131.08, 130.54, 129.43, 46.07, 42.53, 33.81 , 30.78, 20.97, 19.44 ppm.HR–MS(m / z)(ESI): calcd for C 28 H 29 N5O5S2[M+H] + : 580.1693; found: 580.1610.

[0129] 5-(3,5-bis((E)-4-methoxybenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanyl-1,3,4-thiadiazol-2-yl)pentanamide (II14): Yield 66%, as a yellow solid. 1 H NMR (500MHz, DMSO) δ 12.87 (s, 1H), 8.32 (s, 2H), 7.64 (d, J = 13.1 Hz, 2H), 7.51 (t, J =8.4 Hz, 4H), 7.05 (dd, J = 21.9, 8.5 Hz, 4H), 4.81 (d, J = 13.6 Hz, 4H), 3.82(d, J = 7.5 Hz, 6H), 2.44 (t, J = 7.2 Hz, 2H), 2.28 (t, J = 7.1 Hz, 2H), 1.72(p, J = 7.2 Hz, 2H). 13 C NMR (100 MHz, DMSO-d6) δ 185.77, 171.75, 170.38,164.25, 160.96, 160.32, 135.79, 132.49, 130.49, 126.97, 126.69, 114.37,55.31, 46.09, 42.50, 33.82, 30.80, 19.45 ppm.HR–MS (m / z) (ESI): calcd forC 28 H 29 N5O7S2[M+H] + : 612.1587; found: 612.1508.

[0130] 5-(3,5-bis((E)-4-trifluoromethylbenzyl)-4-oxopiperidin-1-yl)-5-oxo-N-(5-sulfanilamide-1,3,4-thiadiazol-2-yl)pentanamide (Ⅱ15): Yield 69%, as a yellow solid. 1 H NMR(500MHz, DMSO) δ 12.87 (s, 1H), 8.31 (s, 2H), 7.89 – 7.71 (m, 10H), 4.84 (s,4H), 2.43 (t, J = 7.3 Hz, 2H), 2.27 (t, J = 7.1 Hz, 2H), 1.70 (p, J = 7.1 Hz, 2H). 13C NMR (100 MHz, DMSO-d6) δ 186.11, 171.74, 170.60, 164.25, 160.96,138.18, 134.52, 130.97, 129.36, 129.11, 125.53, 125.06, 122.89, 45.96,42.24, 33.80, 30.71, 19.36 ppm.HR–MS (m / z) (ESI): calcd for C 28 H 23 F6N5O5S2[M +H] + : 688.1122; found: 688.1045.

[0131] The structural formulas of the target compounds II1 to II15 obtained in this embodiment are shown below:

[0132]

[0133] Example 5: Preparation of target compounds II10, II12, and II15

[0134] Compound II10: Example 4 was repeated, except that acetone was used instead of dichloromethane, TBTU was used instead of HATU, and N,N-diisopropylethylamine was used instead of triethylamine to adjust the pH of the system to 9.5. A yellow solid was finally obtained in 56% yield. Characterization by 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed it to be compound II10.

[0135] Compound II12: Example 4 was repeated, except that triethylamine was not added and HOBT was used instead of HATU. A yellow solid was obtained in 50% yield. Characterization by 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed it to be compound II12.

[0136] Compound II15: Example 4 was repeated, except that tetrahydrofuran was used instead of dichloromethane. A yellow solid was finally obtained in 58% yield. Characterization by 1H NMR, 1C NMR, and high-resolution mass spectrometry confirmed it to be compound II15.

[0137] Example 6: Preparation of single crystals of target compounds II3 and II10

[0138] Compound II3 Single Crystal: 10 mg of Compound II3 prepared in Example 4 was placed in a beaker, and 10 ml of a mixture of acetonitrile and dichloromethane (volume ratio 1:1) was added. Then, 40 ml of water was added, and the beaker was sealed with sealing film and placed in a cool place. After slow evaporation for 5 days, pale yellow needle-like crystals precipitated. Crystals of suitable size were selected for structural analysis using X-ray single-crystal diffraction. Detailed crystal measurement data are shown in Table 2 below. The crystal structure of the obtained crystals is as follows: Figure 4 As shown, the obtained pale yellow needle-like crystals were determined to be single crystals of the target compound II3.

[0139] Compound II10 Single Crystal: 10 mg of Compound II10 prepared in Example 4 was placed in a beaker, and 10 ml of a mixture of acetonitrile and dichloromethane (volume ratio 1:1) was added. Then, 40 ml of water was added, the beaker was sealed with sealing film, and placed in a cool place for slow evaporation for 5 days. Pale yellow triclinic columnar crystals precipitated. Crystals of suitable size were selected for structural analysis using X-ray single-crystal diffraction. Detailed crystal measurement data are shown in Table 2 below. The crystal structure of the obtained crystals is as follows. Figure 5 As shown, the obtained pale yellow triclinic columnar crystals were determined to be single crystals of the target compound II10.

[0140] Table 2: Data on crystallographic and structural corrections for compounds II3 and II10

[0141]

[0142] Experimental Example 1: In vitro antitumor activity assay

[0143] The clinically used drugs doxorubicin hydrochloride, CA inhibitor SLC-0111, and acetazolamide were used as positive control drugs, and the corresponding solvents were used as negative control drugs. Human breast cancer cells MDA-MB-231, human colon cancer cells HCT-116, and SW480 were used as test cell lines. The MTT assay was used to test the in vitro antitumor activity of the target compound of the present invention.

[0144] Cells in good growth condition were seeded at 2×10⁴ to 4×10⁴ cells / well in 96-well plates and cultured at 37 ℃ for 24 h. Then, 20 μL of the test compound (starting concentration 50 μM, serially diluted 2-fold, with 5 replicates per concentration) was added, and the plates were incubated for another 24 h. 20 μL of 5 mg / mL MTT working solution was added to each well, and the plates were incubated for 4 h. The supernatant was carefully discarded, and 150 μL of DMSO was added to dissolve the formazan purple crystals produced by reduction. The absorbance of each well was measured using a microplate reader. The cell proliferation inhibition rate of each well was calculated, and the results are shown in Table 3 below.

[0145] Table 3. Inhibitory activity of the target compounds of this invention against different cell lines.

[0146]

[0147] a IC 50 The values ​​are the mean ± SD of 3 independent experiments.

[0148] As shown in Table 3, some compounds exhibited good antitumor activity. For human breast cancer cells MDA-MB-231, compounds I10, I12, I15, II10, II12, II13, and II15 showed good cytotoxicity (IC50, IC50). 50 The effective values ​​were 7.18±0.97, 6.51±0.70, 4.34±0.28, 4.63±0.01, 4.71±0.56, 6.76±0.9, and 4.21±0.42, respectively. Compounds I15 and II15 were superior to the positive control drug doxorubicin hydrochloride (4.57±0.19). Compounds I10, I12, I15, II10, II12, and II15 showed good cytotoxicity against HCT-116 cells, with IC50 values ​​of 7.18±0.97, 6.51±0.70, 4.34±0.28, 4.63±0.01, 4.71±0.56, 6.76±0.9, and 4.21±0.42. 50 The effective values ​​were 3.12±0.07, 5.18±0.86, 2.54±0.17, 4.41±1.1, 6.22±1.1, and 4.17±1.1, respectively. For SW480 cells, compounds I10, I12, I15, II10, II12, II13, and II15 exhibited good cytotoxicity, with IC50 values ​​of 3.12±0.07, 5.18±0.86, 2.54±0.17, 4.41±1.1, 6.22±1.1, and 4.17±1.1, respectively. 50 The values ​​were 4.17±0.83, 6.74±1.63, 5.26±0.96, 5.31±0.94, 6.23±0.35, 5.73±0.47, and 4.67±0.25, respectively.

[0149] The above in vitro antitumor activity tests show that the sulfonamide-3,5-bis(benzylidene)-4-piperidinone derivative of the present invention is expected to be used in the preparation of antitumor drugs.

[0150] Experiment Example 2: Cell Proliferation Experiment under Hypoxic Environment

[0151] The CA inhibitor SLC-0111 was used as a positive control to simulate hypoxic conditions for cell viability testing, and the following steps were performed:

[0152] MDA-MB-231, HCT-116, and SW480 cells were seeded into 96-well plates at a density of 2×10⁴–4×10⁴ cells / well and incubated at 37°C for 24 h. Fresh culture medium was removed and 100 μM CoCl₂ was added, and the plates were incubated at 37°C for 24 h, followed by CoCl₂ exposure for 48 h. Then, 20 μL of the assay compound (starting concentration 40 μM, 2-fold dilution, 5 replicates per concentration) was added, and the plates were incubated at 37°C for 24 h. Finally, 20 μL of 5 mg / mL MTT working solution was added, and the plates were incubated for 4 h. The supernatant was carefully discarded, and 150 μL of DMSO was added to dissolve the formazan purple crystals produced by reduction. The absorbance of each well was measured using a microplate reader.

[0153] Experimental results are expressed in IC 50 The results are shown in Table 4 below, with SLC-0111 (>50 μM) serving as a positive control. The IC50 values ​​of compounds I10, I12, I15, II10, II12, and II15 on MDA-MB-231 cells under hypoxic conditions are also presented. 50 The IC50 values ​​of compounds I10, I12, I15, II10, II12, and II15 against HCT-116 cells under hypoxic conditions were 4.07±0.64, 4.21±0.33, 2.63±0.12, 3.79±0.87, 4.35±0.53, and 3.38±0.39 μM, respectively, which were all higher than the cytotoxicity values ​​of 7.18±0.97, 6.51±0.70, 4.34±0.28, 4.63±0.01, 4.71±0.56, and 4.21±0.42 μM under normal oxygen conditions. 50 The IC50 values ​​of compounds I10, I12, I15, II10, II12, and II15 against SW480 cells under hypoxic conditions were 1.22±0.11, 1.38±0.39, 0.70±0.21, 1.64±0.24, 3.35±0.52, and 1.86±0.43, respectively, were all higher than those against SW480 cells under normal oxygen conditions (3.12±0.07, 5.18±0.86, 2.54±0.17, 4.41±1.1, 6.22±1.1, and 4.17±1.1, respectively). 50 The cytotoxicity values ​​of 2.26±0.20, 2.54±0.20, 2.81±0.51, 3.16±0.81, 3.54±0.17, and 3.25±0.21 under normal oxygen conditions were all higher than those under normal oxygen conditions (4.17±0.83, 6.74±1.63, 5.26±0.96, 5.31±0.94, 6.23±0.35, and 4.67±0.25). This indicates that the compound further inhibits cell activity under hypoxic conditions by inhibiting CA activity.

[0154] Table 4. Antiproliferative activity of some target compounds of the present invention against MDA-MB-231, HCT-116 and SW480 cell lines under normal oxygen and hypoxic conditions.

[0155]

[0156] a IC 50 The values ​​are the mean ± SD of 3 independent experiments.

[0157] Experimental Example 3: Animal Experiments with Compounds II10, II12, and II15

[0158] The animal experiments for compounds II10, II12, and II15 are as follows:

[0159] The applicant used a xenograft model of breast cancer MDA-MB-231 cells to study the in vivo antitumor activity of compounds II10, II12, and II15. Twenty-four hours after inoculation, mice were randomly divided into nine groups (n=5). Every two days, mice were administered compounds II10 (5 or 10 mg / kg), II12 (5 or 10 mg / kg), II15 (5 or 10 mg / kg), or a carrier via intravenous injection, using the clinical drug doxorubicin hydrochloride (DOX) as a positive control, for 21 consecutive days. Results are as follows... Figure 1 As shown, compared with the control group, compounds II10, II12, and II15 effectively inhibited tumor growth. Figure 1 A), II10 showed tumor growth inhibition rates of 51% and 58% at doses of 5 mg / kg and 10 mg / kg, respectively. Figure 1 B), II12 showed tumor growth inhibition rates of 49% and 61% at doses of 5 mg / kg and 10 mg / kg, respectively. Figure 1 B), II15 showed tumor growth inhibition rates of 52% and 75% at doses of 5 mg / kg and 10 mg / kg, respectively. Figure 1 B). Compared with the blank group, the experimental groups with added compounds II10, II12, and II15 significantly reduced tumor volume (B). Figure 1 C), but the trend of weight change in each group of mice was basically the same as that in the blank group, further demonstrating the safety of compounds II10, II12, and II15. Figure 1 D). In vivo antitumor activity tests in mice showed that compounds II10, II12, and II15 could effectively inhibit the growth of breast cancer MDA-MB-231 tumors.

Claims

1. A sulfadiazine-3,5-bis(benzylidene)-4-piperidinone derivative or a pharmaceutically acceptable salt thereof with the structure shown in formula (I) or formula (II) below: (I), (II); in: R1 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; R2 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; R3 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; n represents 3.

2. The process for the preparation of sulfonamide-3,5-bis(benzylidene)-4- piperidinone derivatives according to claim 1, characterized in that, Take the compound shown in formula (III) below and react it with p-aminobenzenesulfonamide or the compound shown in formula (IV) below in an organic solvent and react it under heating or no heating conditions to obtain the crude product of the compound shown in formula (I) or formula (II); (III), (IV); in: R1 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; R2 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; R3 represents a hydrogen atom, a halogen atom, a C1-C4 alkyl group, a C1-C4 alkoxy group, or a halogenated C1-C4 alkyl group; n represents 3.

3. The preparation method according to claim 2, characterized in that, Add the condensing agent before the reaction.

4. The preparation method according to claim 3, characterized in that, The condensing agent is selected from one or more of HATU, DIPEA, HOBT, EDCI and TBTU.

5. The preparation method according to any one of claims 2 to 4, characterized in that, The reaction takes place under alkaline conditions.

6. The preparation method according to claim 5, characterized in that, The reaction is carried out under alkaline conditions with a pH of 7.5 to 9.

7. The preparation method according to any one of claims 2 to 4, characterized in that, The organic solvent is selected from one or more combinations of acetonitrile, acetone, methanol, ethanol, dichloromethane, chloroform and tetrahydrofuran.

8. The preparation method according to any one of claims 2 to 4, characterized in that, It also includes a step of purifying the crude target compound obtained.

9. Use of the sulfadiazine-3,5-bis(benzylidene)-4-piperidinone derivative of claim 1 or a pharmaceutically acceptable salt thereof in the preparation of a medicament for treating tumors.

10. A pharmaceutical composition comprising a therapeutically effective dose of the sulfadiazine-3,5-bis(benzylidene)-4-piperidinone derivative of claim 1 or a pharmaceutically acceptable salt thereof as an active ingredient, and a pharmaceutically acceptable carrier.

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